HPLC Separation of Caffeine, 3- Methylxanthine, 1- Methylxanthine, Xanthine

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Chromatogram

HPLC Separation of Caffeine, 3- Methylxanthine, 1- Methylxanthine, Xanthine

HPLC Separation of Caffeine, 3- Methylxanthine, 1- Methylxanthine, Xanthine

Description

Application Notes: Xanthines are polar neutral compounds which are hard to retain and separate by traditional reversed-phase chromatography. However a hydrogen bonding method makes separation possible due to an observable correlation between the number of hydrogens available for interaction and retention time. Molecules with no hydrogens available for interactions retain less, and compound with multiple hydrogen donors retain the most. Retention time can be controlled by changing ratio of ACN:MeOH. Other protic and aprotic solvents can be used to control retention time and selectivity of separation.

Application Columns: SHARC 1, 3.2×100 mm, 5 um, 100A, To learn more about SHARC 1 columns click here . To order this column click here . To see more chromatographic separations check our web site.

Application Compounds: Caffeine, 3-methylxanthine, 1-methylxanthine, and xanthine

Method Parameters

Mobile PhaseMeCN/MeOH
BufferAmFm, Formic acid
Flow Rate1.0 ml/min
DetectionUV, 270 nm
Class of CompoundsDrug, Acid, Hydrophilic, Ionizable, Vitamin, Supplements
Analyzing CompoundsCaffeine, 3- Methylxanthine, 1- Methylxanthine, Xanthine

HPLC Column Used

Sharc 1, 3.2×100 mm, 5 µm, 100A